Antimicrobial formulations and methods of using the same

ABSTRACT

Formulations and blends are described comprising zinc, silver, bismuth and copper in non-toxic amounts that are useful as treatments of a wide variety of conditions and diseases promoted by bacteria or other microorganisms.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority under Title 35 United States Code, §119 to U.S. provisional patent application U.S. Pat. App. Ser. No.60/542,636, filed Feb. 18, 2004.

FIELD OF INVENTION

The present invention relates to antimicrobial formulations and methodsof using the same, and more particularly, the present invention relatesto compositions useful to treat bacteria related conditions anddiseases.

BACKGROUND OF THE INVENTION

Copper containing dental cements have been shown to demonstrategermicidal activity. See e.g., M. Dumas and M. Blush. BacteriocidalEffects of Copper Cements: A Review of the Literature, Tufts DentalOutlook, 27 (2): 1-5 (1954). Similarly, silver has also been shown tohave antimicrobial properties. See e.g., Hill et al., RelativeEfficiency of Germicidal Cements, The Journal of the American DentalAssociation, 21 (3): 1565-1571 (1934).

Biofilms are diverse microbial colonies of bacteria, spirochetes, fungi,cocci, viruses, etc. that colonize and begin producing slime(mucopolysaccharides) that makes them invisible to the human immunesystem. Biofilms are resistant to immune cells and antibiotics.Microorganisms in a biofilm survive better and exhibit strongerresistance to the environment than do cells that are not in a biofilm.See e.g, Merritt et al., Bacterial Biofilm and Dentistry, CDA Journal,29 (5): 355-360 (2001)). Biofilms are the chief contributors of dentaldisease. Biofilms can form in various environments, including within themouth and in water supply lines Id.

Microorganisms are responsible for a number of diseases and adverseconditions in mammals. Moreover, while known to be effective againstmicrobial related diseases and conditions, copper at high concentrationsis generally toxic and causes discoloration of the teeth. Hence, thereis a need for antimicrobial formulations that are non-toxic andeffective against a wide variety of organisms and related diseases andconditions.

BRIEF SUMMARY OF THE INVENTION

The present invention is an antimicrobial formulation comprising a zinccompound such as zinc oxide from between about 2 to 90 percent byweight, a silver compound, from between about 005 to 2 percent byweight, a copper compound from between about 0.05 to 10 percent byweight and bismuth between about 2 to 50 percent by weight. The subjectinvention also includes an antimicrobial blend comprising theantimicrobial formulation plus a medium. The medium may be phosphoricacid and water and/or combinations thereof. The formulation of thesubject invention may also be used in combination with a wide variety ofcosmetic and pharmaceutical compositions including skin cream, petroleumjelly, astringent, composite resin, varnish, rose oil, nail polishes,ointments and bonding resins.

The present invention also provides for a novel process for preparingthe antimicrobial formulation and methods of using the antimicrobialformulation to treat a variety of indications or conditions such asbiofilms, skin irritation, and fungal infection.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides for formulations containing low non-toxicamounts of copper, silver, zinc and bismuth that are effective intreating microorganisms found in biofilm and other related diseases andconditions. For dental indications, the formulations of the presentinvention retard biofilm formation, eliminates existing biofilm andpromotes secondary dentin formation. In other applications, indications,diseases or conditions, the formulations of the present invention retardmicrobial growth and treat irritated tissue. Specifically, theantimicrobial formulation can treat the growth of microorganisms ontoenails, fingernails and hooves, and skin irritations. Also, theantimicrobial formulation can be used to treat the contamination ofwater lines. Specifically, the water lines of a dental unit.

The present invention provides formulations and blends that are highlyantimicrobial, non-toxic, biocompatible, non-irritating to dentaltissues, are color-fast, easy to prepare, have adequate setting time,have superior mechanical properties, promote secondary dentin formationand add years of longevity to fillings, castings and endodontic posts.

The formulations of the present invention are useful in combination witha wide variety of dental compositions and treatments including, but notlimited to, resins, fillings, bases, periodontal packs, cements, sealerssuch as root canal sealers or root crack sealers and calcium hydroxidepreparations for direct pulp caps.

The antimicrobial formulation of the subject invention may be providedin a powder or solid form. The formulation may be combined with a mediumsuch as phosphoric acid or water to produce a blend. When theantimicrobial formulation is mixed with phosphoric acid, a zincphosphate cement is made. The formulation may also be mixed with othercompositions and substances described herein and/or used in combinationwith other compositions and substances.

The antimicrobial formulation of the present invention comprises a zinccompound such as zinc oxide from between about 2 to 90 percent byweight, a silver compound, from between about 0.05 to 2 percent byweight, a copper compound from between about 0.05 to 10 percent byweight and bismuth between about 2 to 50 percent by weight. Suitablezinc compounds for use in connection with the present invention includezinc oxide. Silver compounds useful in connection with the subjectinvention include silver chloride, silver phosphate and silver nitrate.Copper compounds useful in connection with the subject invention includecupric oxide, cuprous oxide, cuprous iodide, cupric iodide, cupricphosphate, copper (II) hydrogen phosphate, and cupric silicate.

The antimicrobial formulation of the subject invention may include othercompounds. For example, iron or iron compounds may be added to furtherenhance the antimicrobial effect of the subject formulation. Theantimicrobial formulations and blends of the subject invention may alsobe mixed with other compositions or used in combination with othercompounds or dental compositions. For example, the formulation of thesubject invention may be mixed with calcium hydroxide and placed on atooth to protect against or retard decay. The formulation can also becombined with a composite resin and placed as a base. Also, if a mediumsuch as phosphoric acid is added to the formulation, activated copperions in the zinc phosphate cement leach into the dentin tubules andfurther assist in long term protection against decay and biofilmformation. The formulation of the present invention may also be used incombination with fluoride compounds including but not limited to sodiumfluoride, and stannous fluoride.

The formulations and blends of the subject invention have low non-toxicconcentrations of copper and are effective against biofilm formation aswell as treating existing biofilm. The activated copper ions alsoprotect teeth against decay, dental caries and plaque formation. Hence,the formulation of the subject invention is useful to treat tooth decayalone or in combination with other compounds such as stannous fluorideand sodium fluoride.

Copper is an important constituent of bone, bland, and nerve tissue andregulates the essential balance between all catabolic and anabolicprocesses of tissue metabolism. It is a cofactor in many enzymesnecessary for connective tissue and bone formation, and protects thecells from oxidation as well as destructive affects of toxic agents fromboth external and internal sources. While the presence of ironfacilitates the activity of copper in vital immune functions, copper isindispensable for the synthesis of heme in the heme factor of basalimmunity, and is the prime factor in accelerating the synthesis ofcatalase, an antimicrobial enzyme. Disturbances in copper metabolismcreate nutritive imbalances in the metabolism of bone tissue, andcontribute to build up of toxic effete material, decreased resistance,low immunity, low white blood cell count, degeneration of tissues, andmalignancy. These deficiencies also reduce the life of the red bloodcorpuscles and the capacity of the bone marrow to produce them. Indental indications, copper increases local and general immunity andprohibits chronic inflammation and infection in tissues associated withlocal dental procedures. Copper supports tissue metabolism and enhancingdetoxification of the oral cavity including the teeth and bone tissues.

Iron is another essential component in heme-factor immunity and is alsofound in the oxidation enzymes (cytochromes) of living tissues as wellas in the enzyme myeloperoxidase that is essential for the production ofwhite blood cells. It is also a component of catalase, an intracellularenzyme that destroys toxic byproducts of other metabolic processes andinflammations, as well as being an affective antidote against mineraland heavy metal toxins. Iron is an essential substance for the synthesisof enzymes and immune substances that destroy microorganisms.Disturbances in metabolism of iron result in degenerative processes,chronic infection and inflammation, and low general immunity. In dentalindications, iron increases immune response and is a primary substancein helping to prevent chronic infection and pain in teeth and gums.

The formulations and blends of the present invention may be applied to atooth without the need for the removal of all decayed tissue or exposureof the pulpal tissue. The present invention may be used to promotesecondary dentin formation.

To facilitate the understanding of the invention, a number of terms aredefined below. Terms defined herein have meanings as commonly understoodby a person of ordinary skill in the areas relevant to the presentinvention. Terms such as “a”, “an” and “the” are not intended to referto only a singular entity, but include the general class of which aspecific example may be used for illustration. The terminology herein isused to describe specific embodiments of the invention, but their usagedoes not limit the invention, except as outlined in the claims.

A “biofilm” is a colony of microorganisms attached to a solid surface. Abiofilm colony can include bacteria, spirochetes, fungi, cocci, yeasts,protozoa, and other microorganisms. A “biofilm” includes the slime,mucopolysaccharides and other compounds produced by the microorganismsof a colony.

“Calcination” is the conversion of a metal into its oxide as a result ofheating the metal to a high temperature.

“Dentin” is the calcified tissue surrounding the pulp cavity of a tooth.

“Activated” copper is copper which is missing one or more electrons.

In the formulation of the present invention, the antimicrobial effect ofcopper is increased when it is activated by silver. Silver activatescopper galvanically as silver is a dissimilar metal to copper. Our ionicspin created by the activation of the copper allows the copper ions topenetrate biofilms and to retard microbial growth. This allowsantimicrobial activity to be obtained without use of high concentrationsof copper or silver.

It has been reported that copper ions may be activated by iron.Moreover, activated copper ions have been shown to be useful in certaindental compositions. Bismuth has been used shown to be effective indental compositions containing copper ions. See e.g, M. Dumas and M.Blush. Bacteriocidal Effects of Copper Cements: A Review of theLiterature, Tufts Dental Outlook, 27 (2): 1-5 (1954).

The present formulations may be used in combination with cosmetic and/orpharmaceutical compositions and related compounds such asalkyleneglycols, or alkyleneglycols in combination with one or morederivatives of hydroxyalkylcellulose, alcohols such as ethanol orpropanol, glycols such as butylenes or hexylene glycol, polyols such assorbitol or glycerol, polyethylene or polypropylene glycols.Biologically acceptable hydroxyalkylcelluloses may also be used incombination with the formulation of the present invention.

Topical treatments include use of the antimicrobial blend of the subjectinvention alone or in combination with a tissue compatible vehicle, suchas a lotion-, ointment-, cream- or gel-based vehicle. Such vehicles arewell known in the art and commercially available for formulation ofactive ingredients into a suitable form for topical application.

EXAMPLES Example 1 Antimicrobial Useful as a Treatment for SkinConditions and Disorders

Percentage by weight silica 4 magnesium oxide 6 zinc oxide 81 bismuthsubnitrate 5 cuprous oxide 2 silver chloride 1 titanium oxide 1

Silica, magnesium, zinc oxide and bismuth sub nitrate are first calcined(cinered, calcinated) by mixing the dry powders in sealed mixer. Then,the powder is placed in an airtight porcelain crucible and heated to1,050° C. for three hours. The result is a fine powder, more medicinaland less toxic than an uncalcined powder of the same.

Next, the silver chloride is ground in a low oxygen, low lightenvironment to protect the compound. The silver chloride powder is thenmixed uniformly together with the calcined powder in the sealed mixer.

Cuprous iodide is then added to the mixture and thoroughly mixed in thesealed container: then, titanium oxide is gradually mixed into theformula-starting at 0.25% by weight to a maximum of 1% by weight, untilthe desired shade is achieved. White copper cement is in its untreatedstate, yellowish. Titanium oxide will at 1% make the powder snow white.Different individual preferences for color may thus be accommodated.

This formulation may be used as a powder or mixed with an aqueous ornon-aqueous medium and/or in combination with vaseline jelly and thelike.

Example 2 Antimicrobial Formulation Useful as a White Copper Cement

Percentage by weight silica 4 magnesium oxide 6 zinc oxide 81 bismuthsubnitrate 5 cuprous oxide 2 silver chloride 1 titanium oxide 1

This formulation is antimicrobial, highly biocompatible, color fast,easy to mix, has adequate setting time; easy to clean after setting,adds years of longevity to fillings, castings and endodontic posts andpromotes secondary dentin formation.

This formulation may be used as a powder or mixed with an aqueous ornon-aqueous medium.

As discussed above, the formulation may be heated at high temperaturesfor a prolonged period of time in order to calcinate the white coppercement.

Example 3 Antimicrobial Formulation Useful for Preparing Red CopperCement

Percentage by weight silica 4 magnesium oxide 6 zinc oxide 73 bismuthsubnitrate 5 ferric oxide 3 cupric oxide 7 silver chloride 1

Silica, magnesium, zinc oxide, bismuth sub nitrate and ferric oxide arefirst calcined (cinered) by mixing the dry powders in a sealed mixer.The mixture is then placed in an airtight porcelain crucible and heatedto 1,050° C. for three hours. The result is a fine powder said to bemore medicinal and less toxic than an uncalcined equivalent.

The silver chloride is then made finer by grinding or crushing in a lowoxygen, low light environment to protect the compound from oxidation andcolor change. Following this procedure the silver chloride powder ismixed uniformly along with the above calcined base in the sealed mixer.The cupric oxide may be added at the same point and thoroughly mixed,leaving a red powder which completes the formulation.

This formulation is antimicrobial, highly biocompatible, color fast,easy to mix, has adequate setting time, easy to clean after setting,adds years of longevity to fillings, castings and endodontic posts andpromotes secondary dentin formation. Red copper cement can directlyimpact and reduce tendencies to the pathological patterns and tissues ona bio-energetic functions level including follicular cysts, dentalfistula, dental foot granuloma, fundus abscess, gingival sulcus,ulcerous gingivitis, maxillary ostitis, exudative ostitis, scleroticostitis, pepto-streptococci, borrelia burgdorfer, gangrenous pulpa,acute pulpitis, and caries.

This formulation may be used as a powder or mixed with an aqueous ornon-aqueous medium. This formulation may be heated at high temperaturesfor a prolonged period of time in order to calcinate the formulationused to make red copper cement. The formulation of Example 3 isparticularly useful in preventing and treating dental decay alone or incombination with fluoride compounds such as stanneous fluoride andsodium fluoride between about 0.05 weight percent and 5 weight percent.

Also, the red copper powder of Example 3 may be mixed with a combinationof 400 ppm ethyl ether anhydrous and/or COPALITE WE® (a dental varnishwithout chloroform) to treat fungal toenails. To date, this compositionhas been successful in nine instances in treating fungal toenails thatwere resistant to over the counter and prescription medications.

Example 4 Root Canal Sealer

Percentage by weight zinc oxide 4 partially hydrogenated rosin 25bismuth subcarbonate 14 barium sulfate 11 sodium borate 2 cuprous oxide7 silver chloride 1

This formulation may be mixed in with combination of 400 ppm ethyl etheranhydrous and/or Copalite WE® (a dental varnish without chloroform) andmay be used to seal root canals when eugenol liquid is the medium.

Example 5 Treating Fungal Toenails and Fingernails

Minimum % by weight Maximum % by weight Silver nitrate 0.05 2 Cuprousoxide or 2 10 cupric oxide Bismuth Subnitrate 2 50 Zinc Oxide 2 90Magnesium Oxide 2 10 Echinacea 0.25 5 Antimony 0.05 1 Belladonna 0.050.5

Rose oil is added as an emulsifier in an amount from 1 to 100 grams

The formulation may be used as a powder or mixed with an aqueous ornon-aqueous medium. For example, the formulation may be mixed with anastringent.

The antimicrobial formulation is then applied directly to the afflictedarea as needed.

Example 6 Treating Skin Irritation

Minimum % by weight Maximum % by weight Silver nitrate 0.05 2 Cuprousoxide or 2 10 cupric oxide Bismuth 2 50 Zinc 2 90 Magnesium 2 10Echinacea 0.25 5 Antimony 0.05 1 Belladonna 0.05 0.5

Rose oil is added as an emulsifier in an amount from 1 to 100 grams.

The formulation may be used as a powder or mixed with an aqueous mediumor other composition or substance. For example, the formulation may bemixed with petroleum jelly.

The antimicrobial formulation is then applied directly to the afflictedarea as needed.

Example 7 Treating White Line Disease in Horses

Percentage by weight silica 4 magnesium oxide 6 zinc oxide 81 bismuthsubnitrate 5 cuprous oxide 2 silver chloride 1 titanium oxide 1

White Line Disease in horses is a deterioration of the inner portion ofthe hoof wall and occurs at the hoof wall/sole junction. The separationat the hoof wall/sole junction allows microorganisms to enter.

The formulation may be used as a powder or mixed with an aqueous ornon-aqueous medium or other composition or substance.

The antimicrobial formulation is applied directly to the afflicted areaas needed.

Example 8 Treating Dental Water Lines

Percentage by weight silica 4 magnesium oxide 6 zinc oxide 81 bismuthsubnitrate 5 cuprous oxide 2 silver chloride 1 titanium oxide 1

The formulation may be used as a powder or mixed with an aqueous medium.

The formulation is introduced into the water lines of a dental unit andthe liquid is allowed to remain undisturbed overnight. The next morning,the lines are rinsed for one minute. Commercial test kits indicate thatthe water lines of the dental unit were purged of microbes.

Example 9 Efficacy of Dental Cement in Biofilm Prevention in a StagnantSystem

Three types of standard sized coupons with a fine grained antimicrobialcontaining powder of the dental cement were used (labeled 1, 2 and 3).The tests were performed in stagnant test tubes.

Three types of dental cement coupons (labeled 1, 2 and 3), dental cementpowder and polycarbonate coupons (as an untreated control) were placeinto test tubes with 5 mL sterile tryptic soy media at a concentrationof 30 mg/L. Each type of coupon was placed into 3 separate test tubes(for 3 replicates). The dental cement powder was placed in the test tubea concentration of 1 g/L (5 mg/5 mL). Each of the test tubes was cappedto insure sterility was maintained and allowed to sit for 48 hours tofacilitates dissolution of the biocide from the coated coupons.Following this, each of the test tubes received a 100 μL boluscontaining Staphylococcus aureus at a concentration of 2.8×10³ cells. Asingle control tube for each coupon type (and powder) received noinoculum. Prior to inoculation, S. aureus was grown from frozen stockculture in TSB media to the necessary concentration. Followinginoculation, tubes were covered and incubated for 48 hours, after whichthe coupon was removed and attached biofilm was physically scraped intodilution buffer, disaggregated using a tissue homogenizer and seriallydiluted and plate on R2A agar. Results are reported as CFU/cm² on thecoupon surface.

While uncoated polycarbonate coupons exposed to the S. aureus inoculumindicated cell colonization at an average density of 1.08×10⁶ CFU/cm²,none of the dental cement coupons contained any measurable biofilmaccumulation (Table 1). All dental cement formulations tested, and thedental cement powder, were 100% effective in preventing biofilmaccumulation under the condition tested.

TABLE 1 Coupon CFU/cm² Dental cement #1A (inoculated) 0 Dental cement#1B (uninoculated) 0 Dental cement #1C (inoculated) 0 Dental cement #2A(uninoculated) 0 Dental cement #2B (inoculated) 0 Dental cement #2C(inoculated) 0 Dental cement #3A (inoculated) 0 Dental cement #3B(inoculated) 0 Dental cement #3C (uninoculated) 0 Dental cement #4A(uninoculated) 0 Dental cement #4B (inoculated) 0 Dental cement #4C(inoculated) 0 Polycarbonate #5A (uninoculated) 0 Polycarbonate #5B(inoculated) 5.00E + 05

Example 10 Reactivity Testing and Biological Resonance Testing

The disclosed formulations have been shown by reactivity testing forIgG, IgM and IgA antibody production to be biocompatibility superior toother zinc phosphate cements. Furthermore, biological resonance testinghas shown that the disclosed formulations are less susceptible to thefollowing pathological disorders and/or disease-causing organisms:follicular cysts, dental fistula, dental root granuloma, fundus abscess,gingival sulcus, ulcerous gingivitis, maxillary ostitis, exudativeostitis, sclerotic ostitis, gangrenous pulpa, acute pulpitis, caries,pepto-streptococci and caries.

Example 11 Efficacy of Dental Cement to Prevent Biofilm in a StagnantSystem

The efficacy of the dental cement of the subject invention has beentested in “stagnant” test tubes and shown to be effective in preventingbiofilm formation.

A growth curve study of Streptococcus mutans ATCC 25175 and Lactobacillsparacasei was completed in order to determine optimal mediaconcentration and time necessary for the organisms to reach log phase.S. mutans was grown anaerobically in full-strength Brain Heart Infusion(BHI) broth and L. paracasei was grown aerobically in full strengthLactobacilli MRS broth. Both cultures were incubated at 37° C. Based onthe growth curve study, it was determined that S. mutans reaches logphase after 22 hours of anaerobic incubation in BHI broth at 37° C. L.paracasei reach log phase after 46 hours of aerobic incubationLactobacilli MRS broth at 37° C.

Eight coupons of each type were placed in glass scintillation vials andautoclaved. Sterile BHI broth at a concentration of 37 mg/L and sterileLactobacilli MRS broth at a concentration of 55 mg/L was added toappropriate vials. The tubes were incubated at room temperature for 48hours. The tubes were inoculated with 100 μl of S. mutans or L.paracasei grown for the appropriate amount of time at a concentration10³-10⁴ CFU/ml. Vials inoculated with S. mutans were incubatedanaerobically for 48 hours at room temperature. Vials inoculated with L.paracasei were incubated for 48 hours aerobically at room temperature.

At the end of the 48 hour incubation, the coupons were removed from thevials, scraped to remove biofilm. The biofilm samples were disaggregatedvia homogenization. The samples were diluted and plated on tryptic soyagar. S. mutans samples were incubated anaerobically for 60 hours at 37°C. L. paracasei samples were incubated aerobically for 60 hours at 37°C.

The results of this testing are provided in Table 2 immediately below.

TABLE 2 Test Tube Designation, Inoculum, Media and Average Colonization.Tube Coupon Average Colonization Numbers Type Innoculum Media (CFU/cm²)1-3 GLS S. mutans BHI 0 4-6 GLS Lactobacillus MRS 684  7 GLS none BHI 0 8 GLS none MRS 0  9-11 RCC S. mutans BHI 0 12-14 RCC Lactobacillus MRS0 15 RCC none BHI 0 16 RCC none MRS 0 17-19 WC S. mutans BHI 0 20-22 WCLactobacillus MRS 0 23 WC none BHI 0 24 WC none MRS 0 25-27 WC+ S.mutans BHI 0 28-30 WC+ Lactobacillus MRS 0 31 WC+ none BHI 0 32 WC+ noneMRS 0 33-35 PC S. mutans BHI 0 36-38 PC Lactobacillus MRS 210 39 PC noneBHI 0 40 PC none MRS 0Cement Coupons:GLS=Fuji II Glass Ionomer Cement Lot#75 N05K K by GC AmericaRCC=DOC'S BEST Red Copper Zinc Phosphate Cement (Lot# A2920-9)WC=DOC'S BEST White Copper Zinc Phosphate Cement (Lot#i3810-6)WC+=White Copper of Example 2PC=Durelon Cement Lot#166317 by 3M

RCC is made from a formulation of the following compounds in theapproximate amounts by weight percent: zinc oxide (75-85); magnesiumoxide (7-8.5); cupric oxide (5.5-6.5); bismuth subnitrate (2.5-3.5);silica (1.5-2.5); iron oxide (less than 1.5); and sodium fluoride (lessthan 0.1) and mixed with phosphoric acid medium.

WC is made from a formulation of the following in the approximateamounts by weight percent: zinc oxide (75-85); magnesium oxide (7-8.5);cuprous oxide (6.5-7.5); bismuth subnitrate (2.5-3.5); silica (1.5-2.5);and sodium fluoride (less than 0.1) and mixed with a phosphoric acidmedium.

Although the invention has been described with reference to specificembodiments, these descriptions are not meant to be construed in alimiting sense. Various modifications of the disclosed embodiments, aswell as alternative embodiments of the invention will become apparent topersons skilled in the art upon reference to the description of theinvention. It should be appreciated by those skilled in the art that theconception and the specific embodiment disclosed may be readily utilizedas a basis for modifying or designing other structures for carrying outthe same purposes of the present invention. It should also be realizedby those skilled in the art that such equivalent constructions do notdepart from the spirit and scope of the invention as set forth in theappended claims.

It is therefore, contemplated that the claims will cover any suchmodifications or embodiments that fall within the true scope of theinvention.

1. A formulation comprising: (a) about 4% by weight silica; (b) about 6%by weight magnesium oxide; (c) about 73% by weight zinc oxide; (d) about5% by weight bismuth subnitrate; (e) about 3% by weight ferric oxide;(f) about 7% by weight cupric oxide; and (g) about 1% by weight silverchloride.
 2. A formulation comprising: (a) about 4% by weight silica;(b) about 6% by weight magnesium oxide; (c) about 81% by weight zincoxide; (d) about 5% by weight bismuth subnitrate; (e) about 2% by weightcupric oxide; (f) about 1% by weight silver chloride; and (g) about 1%by weight titanium chloride.
 3. A formulation comprising: (a) about 4%by weight zinc oxide; (b) about 25% by weight staybelite resin; (c)about 14% by weight bismuth subcarbonate; (d) about 11% by weight bariumsulfate; (e) about 2% by weight sodium borate; (f) about 7% by weightcuprous oxide; and (g) about 1% by weight silver chloride.
 4. A kitcomprising: (1) an antimicrobial formulation comprising: (a) about 4% byweight silica;(b) about 6% by weight magnesium oxide; (c) about 73% byweight zinc oxide; (d) about 5% by weight bismuth subnitrate; (e) about3% by weight ferric oxide; (f) about 7% by weight cupric oxide; and (g)about 1% by weight silver chloride; and (2) phosphoric acid, a dentalvarnish, stannous fluoride or sodium fluoride.
 5. A kit comprising: (1)an antimicrobial formulation comprising: (a) about 4% by weight silica;(b) about 6% by weight magnesium oxide; (c) about 81% by weight zincoxide; (d) about 5% by weight bismuth subnitrate; (e) about 2% by weightcupric oxide; (t) about 1% by weight silver chloride; and (g) about 1%by weight titanium chloride; and (2) phosphoric acid, a dental varnish,stannous fluoride or sodium fluoride.
 6. A kit comprising: (1) anantimicrobial formulation comprising: (a) about 4% by weight zinc oxide;(b) about 25% by weight staybelite resin; (c) about 14% by weightbismuth subcarbonate; (d) about 11% by weight barium sulfate; (e) about2% by weight sodium borate;(f) about 7% by weight cuprous oxide; and (g)about 1% by weight silver chloride; and (2) phosphoric acid, a dentalvarnish, stannous fluoride or sodium fluoride.